The JI PBL Intereron Source
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     
 

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zheng, H.
Right arrow Articles by Li, Z.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zheng, H.
Right arrow Articles by Li, Z.
The Journal of Immunology, 2001, 167: 6731-6735.
Copyright © 2001 by The American Association of Immunologists

Cell Surface Targeting of Heat Shock Protein gp96 Induces Dendritic Cell Maturation and Antitumor Immunity1

Hong Zheng2, Jie Dai2, Diliana Stoilova and Zihai Li3

Center for Immunotherapy of Cancer and Infectious Diseases, University of Connecticut School of Medicine, Farmington, CT 06030


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results and Discussion
 References
 
gp96 is a residential heat shock protein of the endoplasmic reticulum that has been implicated in the activation of dendritic cells (DCs) for the initiation of adaptive immunity. By genetic targeting of gp96 onto the cell surface, we demonstrate that direct access of gp96 to DCs induces their maturation, resulting in secretion of proinflammatory cytokines IL-1{beta}, IL-12, and chemokine monocyte chemoattractant protein-1 and up-regulation of the expression of MHC class I, MHC class II, CD80, CD86, and CD40. Furthermore, surface expression of gp96 on tumor cells renders them regressive via a T lymphocyte-dependent mechanism. This work reinforces the notion that gp96 is an endogenous DC activator and unveils that the context in which Ag is delivered to the immune system, in this case surface expression of gp96, has profound influence on immunity. It also establishes a principle of bridging innate and adaptive immunity for cancer immunotherapy by surface targeting of an intracellular heat shock protein.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results and Discussion
 References
 
The context in which Ags are delivered to the immune system plays an important role in the initiation of the adaptive immune response (1, 2). It is now clear that antigenic stimulation alone (signal 1) is insufficient for the induction of productive immunity as characterized by priming and expansion of naive T or B lymphocytes (3). Such a process is also critically dependent on the engagement of lymphocytes with a family of costimulatory molecules (signal 2) delivered by the activated dendritic cells (DCs).4 Therefore, the proper context for generating an Ag-specific immune response is when Ag delivery coincides with the activation of DCs (4). Two broad classes of DC activators have been unveiled: exogenous molecules typified by bacterial product LPS (5) and endogenous activators, including the ubiquitously present intracellular heat shock proteins (HSPs) or stress proteins (6, 7, 8, 9, 10, 11, 12, 13) and type I IFN induced after virus infection (14, 15).

gp96 is a prototypical HSP that can bind to the surface of APCs in a receptor-dependent manner (16, 17, 18). One of its receptors has been identified as CD91 (19). The binding of gp96 to its receptor(s) induces DC maturation (11, 12, 13) and facilitates the transfer of gp96-associated peptides from the extracellular compartment to MHC class I molecules for the recognition by Ag-specific CD8+ T cells (19, 20, 21). gp96 resides normally in the lumen of the endoplasmic reticulum (ER) (22, 23, 24). Because of the above immunological properties of gp96 and the importance of the context of Ag delivery on immunity, we reasoned that surface targeting of gp96 on tumor cells would have profound immunological consequences. In this study, we have indeed conducted such directed surface expression of gp96 by genetic and cellular engineering, and we report that it has a dramatic effect on both innate and adaptive immunity. We show that direct access of gp96 to the immune system induces robust maturation of DCs in vitro and primes tumor-specific T cells in vivo. Our data lay further support for the notion that gp96 is one of the endogenous DC activators. Strikingly, it reveals the importance of the context during which Ag is delivered, in this case cell surface expression of HSP gp96, in the initiation of antitumor immune responses.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results and Discussion
 References
 
Mice, cell lines, and Abs

All mice were obtained from The Jackson Laboratory (Bar Harbor, ME) and were maintained by the Center for Laboratory Animal Care at the University of Connecticut Health Center (Farmington, CT) using standard guidelines. Parental tumor cell lines Meth A and CT-26 were obtained from Dr. P. K. Srivastava (University of Connecticut School of Medicine, Farmington, CT). Abs were from BD PharMingen (San Diego, CA) except where indicated.

Construction of vectors for surface expression of gp96

The KDEL-minus murine gp96 cDNA was obtained by RT-PCR of mRNA isolated from normal BALB/c mouse liver and cloned into an EcoRI and SacII double-digested pDisplay vector (Invitrogen, Carlsbad, CA). The hemmaglutinin tag was removed from the original vector. The sequence of the construct was verified.

Transfection and confocal microscopy

Meth A and CT-26 were transfected by electroporation and LipofectAMINE (Life Technologies, Carlsbad, CA), respectively, and selected by G418. gp96 surface-expressing cells were identified by FACS using Abs against gp96 (Charles River Breeding Laboratories, Wilmington, MA) or myc tag (Invitrogen) and were subcloned by limiting dilution. Surface expression was also confirmed by confocal microscopy after fixation and staining with rabbit anti-gp96 Ab, followed by an Alexa Fluor 594-conjugated anti-rabbit IgG (Molecular Probes, Eugene, OR) and costaining with FITC-conjugated anti-Kd mAb. Optical sections were obtained by laser confocal microscopy using an LSM 410 microscope (Zeiss, Oberkochen, Germany).

DC culture, in vitro maturation assay

Syngeneic bone marrow-derived immature DCs were prepared after 6-day culture in the presence of GM-CSF (20 ng/ml; BD PharMingen) following the published protocol (22). Typically, 5 x 105 day +6 immature DCs were incubated with LPS (0.1 µg/ml), or 2 x 105 live tumor cells with or without cell surface gp96 in the presence or absence of polymyxin B (10 µg/ml; Sigma-Aldrich, St. Louis, MO) for 20 h in a 12-well plate. In some experiments, tumor cells and DCs were separated by a 0.4-µm Trans-well filter (Millipore, Bedford, MA). Cytokine production was measured by ELISA kits (monocyte chemoattractant protein-1 (MCP-1) from R&D Systems, Minneapolis, MN; IL-1{beta} and IL-12 from Endogen, Woburn, MA).

Tumor rejection assay

The immunogenicity of tumor cells was tested by inoculating them intradermally into the naive syngeneic mice and athymic nude mice. Tumor growth was monitored twice a week and recorded using vernier calipers, measuring both the longitudinal and the transverse diameters. Average diameters (mm) of the two axes were plotted. For the cross-protection experiment, 5 x 105 Meth A-96tm or Meth A-neo was injected into the left flank, and 1 x 105 Meth A-WT was injected in the right flank. In some experiments, mice were injected with a mixture of Meth A-96tm (5 x 105) and Meth A-WT (1 x 105). Tumor growth kinetics was compared with Meth A-WT alone. There were five mice in each group.

ELISPOT

BALB/c mice were immunized intradermally with 5 x 105 live tumor cells with or without surface gp96 expression. Seven days after the immunization, the total splenocytes (2 x 106 cells/well) were obtained and incubated with irradiated (100 Gy) Meth A-WT (5 x 104 cells/well) in 96-well IP-Multiscreen plates (Millipore) precoated with 100 µl of mouse anti-mouse IFN-{gamma} Ab (10 µg/ml, clone R4–6A2; BD PharMingen). This was followed sequentially by washing, incubation with biotinylated anti-IFN-{gamma} (XMG1.2) mAb, and an HRP-based secondary Ab (Vector Laboratories, Burlingame, CA). The spots were developed with 3-amino-9-ethylcarbazole and H2O2 (Sigma-Aldrich), counted with a Zeiss ELISPOT reader, and reported as the number of IFN-{gamma} spots per 1 x 106 cells.


   Results and Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
References
 
Surface expression of gp96

gp96 resides normally in the lumen of ER due to the presence of an ER retention signal, KDEL, at its carboxyl terminus. In stress and other conditions, both secretion (23) and surface expression of gp96 have been reported (24, 25). To study the direct impact of extracellular gp96 on both innate and adaptive immunity, we targeted gp96 onto the cell surface of Meth A fibrosarcoma and CT-26 colon carcinoma. If gp96 indeed possesses intrinsic immunological properties, we reasoned that cell surface expression of gp96 would have measurable immunological consequences. The construction of the expression vector for cell surface gp96 (96tm) was achieved by deletion of the KDEL sequence followed sequentially by fusion in frame to a c-myc tag and a transmembrane domain from platelet-derived growth factor receptor (Fig. 1GoA). The surface expression was confirmed by confocal microscopy (Fig. 1GoB) and flow cytometry (Fig. 1GoC) using Abs specific for gp96 and the myc tag. Only tumor cells that were transfected for 96tm, but not the control cells transfected with the empty vector (neo), expressed consistently high levels of gp96 on the cell surface. Surface staining by anti-myc Ab proved that the 96tm is expressed as a type I transmembrane protein. Because the cytoplasmic tail retained from the platelet-derived growth factor receptor is only five amino acids in length, it is extremely unlikely that 96tm has any direct signaling property.



View larger version (33K):
[in this window]
[in a new window]
  		FIGURE 1. Expression of gp96 on the cell surface. A, Schematic representation of wild-type gp96 and 96tm protein domain organization. SP, Signal peptide. B, Confocal fluorescence microscopy after staining with anti-gp96 Ab and secondary Ab conjugated with Alexa Fluor 594, followed by costaining with FITC-conjugated mAb against MHC class I (kd). C, FACS analysis of surface expression of gp96 after staining with an irrelevant Ab (green) and Abs against myc tag (red) and gp96 (black).

 
Gp96 surface expression leads to activation of DCs in vitro

We next examined whether the presence of surface gp96 allows tumor cells to mature DCs. Murine bone marrow-derived immature DCs were cocultured for 20 h with tumor cells with or without cell surface expression of gp96. The 96tm-expressing tumor cells, but not control transfectants, activated DCs efficiently as evidenced by up-regulation of costimulatory molecules CD40, CD80, and CD86 as well as MHC class I and MHC class II (Fig. 2GoA). Furthermore, we found that the activation of DCs by 96tm, but not neo transfectants, occurred even when cells had undergone radiation-induced apoptosis (data not shown), indicating that the effect was not dependent on active proliferation of 96tm-expressing cells. All culture media were endotoxin tested to be free of LPS. As expected, DC maturation was not blocked by addition of polymyxin B, which inhibited the effect of LPS completely (Fig. 2GoB). To rule out possible effect from other diffusible agents such as mycoplasma, cell debris, or DNA fragments, DCs and tumor cells were separated by a 0.4-µm Trans-well filter during coculturing. The maturation effect was abrogated (Fig. 2GoB), indicating a requirement for direct contact between DCs and 96tm-expressing tumor cells. In addition, matured DCs secreted proinflammatory cytokines IL-12, IL-1{beta}, and chemokine MCP-1 after coculturing with 96tm (Fig. 2GoC). The profile of cytokine induction by LPS and 96tm is clearly different.



View larger version (45K):
[in this window]
[in a new window]
  		FIGURE 2. Maturation of bone marrow-derived DCs by gp96 surface-expressing tumor cells. A, Murine immature DCs were cultured with medium alone, or tumor cells transfected with 96tm or neo for 20 h were analyzed by flow cytometry. The percentages of CD11C+ cells that were also high for MHC class I, MHC class II, CD80, CD86, and CD40 expression are indicated in the upper right quadrant. Shown here are representative data from numerous experiments using CT-26 transfectants. Meth A-96tm activated DCs with a similar pattern (data not shown). B and C, Immature DCs were cultured with medium alone, LPS, neo, or 96tm for 20 h in the presence () or absence ({square}) of polymyxin B. As a control, tumor cells were also separated from DCs by a Trans-well membrane ({blacksquare}) during some coculture experiments, as indicated. Mature DCs expressing both CD86 and CD11c were analyzed by flow cytometry (B). The supernatants were measured for the secretions of IL-12, IL-1{beta}, and MCP-1 by the respective ELISAs (C).

 
DC maturation as a result of surface expression of gp96 was confirmed in both Meth A and CT-26 using multiple transfectants. Furthermore, using a nonreplicating adenovirus vector, we found that direct expression of 96tm on DCs, but not a control protein {beta}-galactosidase, led to DC activation (data not shown). Our data do not exclude the possibility that other molecules, as a result of gp96 surface expression, are involved in the activation of DCs. Formal proof awaits detailed structure-function analysis of gp96 and the availability of conformation-specific Ab or compound against the receptor-binding site of gp96. Nevertheless, our result, in conjunction with the findings that soluble gp96 activates DCs in vitro (11, 13), matures and promotes DC trafficking to the draining lymph nodes in vivo (12), argues strongly for the idea that gp96 itself is a strong DC activator. Moreover, we showed that the induction of DC maturation occurred regardless of whether gp96 surface-expressing cells were live or apoptotic. Thus, DC activation reported by others as being induced by necrotic but not apoptotic cell lysate (15, 26) is perhaps not so much due to the mode of cell death, but instead is likely dependent upon whether HSPs are accessible to DCs. This explanation is consistent with the finding that HSPs are released when cells undergo necrosis, but not apoptosis (11).

Surface targeting of gp96 increases immunogenicity of tumor cells

We next studied the immunogenicity of gp96 surface-expressing tumor cells. We hypothesized that direct access of gp96 to the immune system via surface expression on tumor cells facilitates the interaction of tumor cells with DCs, leading to enhanced priming of tumor-specific T cells. Meth A-WT grows progressively in BALB/c mice. Intradermal injection of 1 x 105 live Meth A-WT or Meth A-neo led to progressive tumor growth in all the mice. By contrast, injection of up to 5 x 105 live gp96 Meth A-96tm induced efficient T cell-mediated tumor rejection (Fig. 3Go, A and B). By depletion of various cellular components, we found that tumor-specific protection is dependent on CD8+ cells (data not shown). Furthermore, a single immunization with Meth A-96tm, but not with Meth A-WT or Meth A-neo significantly induced the expansion of Meth A-specific IFN-{gamma}-producing T cells without any further in vitro stimulation (Fig. 3GoE). We also demonstrated that the immunity elicited by Meth A-96tm is cross-protective against the parental Meth A. The injection of Meth A-96tm mixed with Meth A-WT led to rejection of both, in contrast to progressive growth kinetics of Meth A-WT alone (Fig. 3GoC). Inoculation of Meth A-96tm to the left flank and Meth A-WT to the right flank of the animals resulted in regression of not only Meth A-96tm, but also the contralateral Meth A-WT (Fig. 3GoD). The successful rejection of parental tumor cells both proximally and distally as a result of immunization with gp96 surface-expressing tumors demonstrated that the 96tm gene targeting approach has therapeutic potential.



View larger version (29K):
[in this window]
[in a new window]
  		FIGURE 3. Surface expression of gp96 on tumor cells leads to tumor rejection and enhanced T cell priming. A, Rejection of Meth A-96tm (•) but not Meth A-neo ({circ}) by wild-type BALB/c mice. B, Progressive growth of both Meth A-96tm (•) and Meth A-neo ({circ}) in nu/nu BALB/c mice. C, Injection of the mixture of Meth A-96tm with Meth A-WT (•) led to rejection of both, compared with progressive tumor growth of Meth A-WT mixed with Meth A-neo ({circ}). D, Simultaneous injection of Meth A-96tm (•) but not Meth A-neo ({circ}) to the left flank of the mice resulted in rejection of Meth A-WT injected at the right flank of the same mice. E, Day 7 splenocytes from mice immunized with PBS, Meth A-WT, Meth A-neo, and Meth A-96tm were assayed directly ex vivo for the frequency of IFN-{gamma}-producing cells by ELISPOT in response to media, Meth A, or CT-26 without further in vitro stimulations. Three independent experiments were performed with similar results.

 
Thus, we have demonstrated that forced surface expression of gp96 on tumor cells stimulates DC maturation in vitro and induces efficient T cell priming and tumor rejection in vivo. Because gp96 can be displayed on the cell surface during stress and under certain physiological conditions (23, 24, 25, 27), it is perhaps reasonable to speculate about the immunological significance of surface gp96. We would like to suggest that the extracellular targeting (secretion and cell surface expression) of intracellular HSPs such as gp96 have physiological bearings (Fig. 4Go). Indeed, it was found that tumor cells secreting a gp96-Ig fusion protein had increased immunogenicity (28). We are actively studying the basis for the dynamics and plasticity of subcellular localization of gp96 and the influences of "ectopic" trafficking of gp96 on immunity in both physiological and pathophysiological conditions.



View larger version (43K):
[in this window]
[in a new window]
  		FIGURE 4. A model for the roles of cell surface gp96 in immunity. Surface expression of gp96 leads to maturation of DCs, as supported by this work (shaded). Soluble gp96 can also be released after pathological cell death. gp96 in this fashion serves essentially as an endogenous adjuvant.

 
Our finding is the first report of the use of HSPs targeted onto the cell surface for potential cancer immunotherapy. Our data, together with those of others (28, 29), should stimulate a new wave of experiments in designing or discovering pharmacological agents that promote HSP export for cancer therapy in the future.


   Acknowledgments
 
We are grateful to E. R. Podack (University of Miami, Miami, FL) for stimulating discussions during the early course of this work. We thank P. K. Srivastava for advice and a critical reading of the manuscript.


   Footnotes
 
1 This work was supported in part by National Institutes of Health Grant CA90337 (to Z.L.). Back

2 H.Z. and J.D. contributed equally to this work. Back

3 Address correspondence and reprint requests to Dr. Zihai Li, Center for Immunotherapy of Cancer and Infectious Diseases, University of Connecticut School of Medicine, MC1601, 263 Farmington Avenue, Farmington, CT 06030-1601. E-mail address: zli{at}up.uchc.edu Back

4 Abbreviations used in this paper: DC, dendritic cell; HSP, heat shock protein; ER, endoplasmic reticulum; KDEL, Lys-Asp-Glu-Leu; MCP-1, monocyte chemoattractant protein-1. Back

Received for publication August 17, 2001. Accepted for publication October 9, 2001.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 References
 

  1. Jr Janeway, C. A.. 1989. Approaching the asymptote? Evolution and revolution in immunology. Cold Spring Harbor Symp. Quant. Biol. 54:1.
  2. Matzinger, P.. 1994. Tolerance, danger, and the extended family. Annu. Rev. Immunol. 12:991.[Medline]
  3. Bretscher, P., M. Cohn. 1970. A theory of self-nonself discrimination. Science 169:1042.[Abstract/Free Full Text]
  4. Banchereau, J., R. M. Steinman. 1998. Dendritic cells and the control of immunity. Nature 392:245.[Medline]
  5. De Smedt, T., B. Pajak, E. Muraille, L. Lespagnard, E. Heinen, P. De Baetselier, J. Urbain, O. Leo, M. Moser. 1996. Regulation of dendritic cell numbers and maturation by lipopolysaccharide in vivo. J. Exp. Med. 184:1413.[Abstract/Free Full Text]
  6. Kol, A., A. H. Lichtman, R. W. Finberg, P. Libby, E. A. Kurt-Jones. 2000. Cutting edge: heat shock protein (HSP) 60 activates the innate immune response: CD14 is an essential receptor for HSP60 activation of mononuclear cells. J. Immunol. 164:13.[Abstract/Free Full Text]
  7. Cho, B. K., D. Palliser, E. Guillen, J. Wisniewski, R. A. Young, J. Chen, H. N. Eisen. 2000. A proposed mechanism for the induction of cytotoxic T lymphocyte production by heat shock fusion proteins. Immunity 12:263.[Medline]
  8. Asea, A., S. K. Kraeft, E. A. Kurt-Jones, M. A. Stevenson, L. B. Chen, R. W. Finberg, G. C. Koo, S. K. Calderwood. 2000. HSP70 stimulates cytokine production through a CD14-dependant pathway, demonstrating its dual role as a chaperone and cytokine. Nat. Med. 6:435.[Medline]
  9. Moroi, Y., M. Mayhew, J. Trcka, M. H. Hoe, Y. Takechi, F. U. Hartl, J. E. Rothman, A. N. Houghton. 2000. Induction of cellular immunity by immunization with novel hybrid peptides complexed to heat shock protein 70. Proc. Natl. Acad. Sci. USA 97:3485.[Abstract/Free Full Text]
  10. Castellino, F., P. E. Boucher, K. Eichelberg, M. Mayhew, J. E. Rothman, A. N. Houghton, R. N. Germain. 2000. Receptor-mediated uptake of antigen/heat shock protein complexes results in major histocompatibility complex class I antigen presentation via two distinct processing pathways. J. Exp. Med. 191:1957.[Abstract/Free Full Text]
  11. Basu, S., R. J. Binder, R. Suto, K. M. Anderson, P. K. Srivastava. 2000. Necrotic but not apoptotic cell death releases heat shock proteins, which deliver a partial maturation signal to dendritic cells and activate the NF-{kappa}B pathway. Int. Immunol. 12:1539.[Abstract/Free Full Text]
  12. Binder, R. J., K. M. Anderson, S. Basu, P. K. Srivastava. 2000. Cutting edge: heat shock protein gp96 induces maturation and migration of CD11c+ cells in vivo. J. Immunol. 165:6029.[Abstract/Free Full Text]
  13. Singh-Jasuja, H., H. U. Scherer, N. Hilf, D. Arnold-Schild, H. G. Rammensee, R. E. Toes, H. Schild. 2000. The heat shock protein gp96 induces maturation of dendritic cells and down-regulation of its receptor. Eur. J. Immunol. 30:2211.[Medline]
  14. Luft, T., K. C. Pang, E. Thomas, P. Hertzog, D. N. Hart, J. Trapani, J. Cebon. 1998. Type I IFNs enhance the terminal differentiation of dendritic cells. J. Immunol. 161:1947.[Abstract/Free Full Text]
  15. Gallucci, S., M. Lolkema, P. Matzinger. 1999. Natural adjuvants: endogenous activators of dendritic cells. Nat. Med. 5:1249.[Medline]
  16. Arnold-Schild, D., D. Hanau, D. Spehner, C. Schmid, H. G. Rammensee, H. de la Salle, H. Schild. 1999. Cutting edge: receptor-mediated endocytosis of heat shock proteins by professional antigen-presenting cells. J. Immunol. 162:3757.[Abstract/Free Full Text]
  17. Wassenberg, J. J., C. Dezfulian, C. V. Nicchitta. 1999. Receptor mediated and fluid phase pathways for internalization of the ER Hsp90 chaperone GRP94 in murine macrophages. J. Cell. Sci. 112:2167.[Abstract]
  18. Binder, R. J., M. L. Harris, A. Menoret, P. K. Srivastava. 2000. Saturation, competition, and specificity in interaction of heat shock proteins (hsp) gp96, hsp90, and hsp70 with CD11b+ cells. J. Immunol. 165:2582.[Abstract/Free Full Text]
  19. Binder, R. J., D. K. Han, P. K. Srivastava. 2000. CD91: a receptor for heat shock protein gp96. Nat. Immunol. 1:151.[Medline]
  20. Basu, S., R. J. Binder, T. Ramalingam, P. K. Srivastava. 2001. CD91 is a common receptor for heat shock proteins gp96, hsp90, hsp70, and calreticulin. Immunity 14:303.[Medline]
  21. Singh-Jasuja, H., R. E. Toes, P. Spee, C. Munz, N. Hilf, S. P. Schoenberger, P. Ricciardi-Castagnoli, J. Neefjes, H. G. Rammensee, D. Arnold-Schild, H. Schild. 2000. Cross-presentation of glycoprotein 96-associated antigens on major histocompatibility complex class I molecules requires receptor-mediated endocytosis. J. Exp. Med. 191:1965.[Abstract/Free Full Text]
  22. Lutz, M. B., N. Kukutsch, A. L. Ogilvie, S. Rossner, F. Koch, N. Romani, G. Schuler. 1999. An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J. Immunol. Methods 223:77.[Medline]
  23. Booth, C., G. L. Koch. 1989. Perturbation of cellular calcium induces secretion of luminal ER proteins. Cell 59:729.[Medline]
  24. Altmeyer, A., R. G. Maki, A. M. Feldweg, M. Heike, V. P. Protopopov, S. K. Masur, P. K. Srivastava. 1996. Tumor-specific cell surface expression of the KDEL containing endoplasmic reticular heat shock protein gp96. Int. J. Cancer 69:340.[Medline]
  25. Wiest, D. L., A. Bhandoola, J. Punt, G. Kreibich, D. McKean, A. Singer. 1997. Incomplete endoplasmic reticulum (ER) retention in immature thymocytes as revealed by surface expression of "ER-resident" molecular chaperones. Proc. Natl. Acad. Sci. USA 94:1884.[Abstract/Free Full Text]
  26. Sauter, B., M. L. Albert, L. Francisco, M. Larsson, S. Somersan, N. Bhardwaj. 2000. Consequences of cell death: exposure to necrotic tumor cells, but not primary tissue cells or apoptotic cells, induces the maturation of immunostimulatory dendritic cells. J. Exp. Med. 191:423.[Abstract/Free Full Text]
  27. Seddiki, N., F. Nato, P. Lafaye, Z. Amoura, J. C. Piette, J. C. Mazie. 2001. Calreticulin, a potential cell surface receptor involved in cell penetration of anti-dna antibodies. J. Immunol. 166:6423.[Abstract/Free Full Text]
  28. Yamazaki, K., T. Nguyen, E. R. Podack. 1999. Cutting edge: tumor secreted heat shock-fusion protein elicits CD8 cells for rejection. J. Immunol. 163:5178.[Abstract/Free Full Text]
  29. Multhoff, G., C. Botzler, L. Jennen, J. Schmidt, J. Ellwart, R. Issels. 1997. Heat shock protein 72 on tumor cells: a recognition structure for natural killer cells. J. Immunol. 158:4341.[Abstract]



This article has been cited by other articles:


Home page
 Int ImmunolHome page
A. Osterloh, A. Veit, A. Gessner, B. Fleischer, and M. Breloer
Hsp60-mediated T cell stimulation is independent of TLR4 and IL-12
Int. Immunol., March 1, 2008; 20(3): 433 - 443.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
R. M. Srivastava, C. Varalakshmi, and A. Khar
The Ischemia-Responsive Protein 94 (Irp94) Activates Dendritic Cells through NK Cell Receptor Protein-2/NK Group 2 Member D (NKR-P2/NKG2D) Leading to Their Maturation
J. Immunol., January 15, 2008; 180(2): 1117 - 1130.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurosci.Home page
M. W. Graner, R. I. Cumming, and D. D. Bigner
The Heat Shock Response and Chaperones/Heat Shock Proteins in Brain Tumors: Surface Expression, Release, and Possible Immune Consequences
J. Neurosci., October 17, 2007; 27(42): 11214 - 11227.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Pathol.Home page
J. M. Han, S. G. Park, B. Liu, B.-J. Park, J. Y. Kim, C. H. Jin, Y. W. Song, Z. Li, and S. Kim
Aminoacyl-tRNA Synthetase-Interacting Multifunctional Protein 1/p43 Controls Endoplasmic Reticulum Retention of Heat Shock Protein gp96: Its Pathological Implications in Lupus-Like Autoimmune Diseases
Am. J. Pathol., June 1, 2007; 170(6): 2042 - 2054.
[Abstract] [Full Text] [PDF]

Home page
 Cardiovasc ResHome page
A. Shamaei-Tousi, J. P. Halcox, and B. Henderson
Stressing the obvious? Cell stress and cell stress proteins in cardiovascular disease
Cardiovasc Res, April 1, 2007; 74(1): 19 - 28.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
B. Liu, Y. Yang, J. Dai, R. Medzhitov, M. A. Freudenberg, P. L. Zhang, and Z. Li
TLR4 Up-Regulation at Protein or Gene Level Is Pathogenic for Lupus-Like Autoimmune Disease
J. Immunol., November 15, 2006; 177(10): 6880 - 6888.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Warger, N. Hilf, G. Rechtsteiner, P. Haselmayer, D. M. Carrick, H. Jonuleit, P. von Landenberg, H.-G. Rammensee, C. V. Nicchitta, M. P. Radsak, et al.
Interaction of TLR2 and TLR4 Ligands with the N-terminal Domain of Gp96 Amplifies Innate and Adaptive Immune Responses
J. Biol. Chem., August 11, 2006; 281(32): 22545 - 22553.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
X.-Y. Wang, H. Arnouk, X. Chen, L. Kazim, E. A. Repasky, and J. R. Subjeck
Extracellular Targeting of Endoplasmic Reticulum Chaperone Glucose-Regulated Protein 170 Enhances Tumor Immunity to a Poorly Immunogenic Melanoma
J. Immunol., August 1, 2006; 177(3): 1543 - 1551.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
N. C. Di Paolo, S. Tuve, S. Ni, K. E. Hellstrom, I. Hellstrom, and A. Lieber
Effect of Adenovirus-Mediated Heat Shock Protein Expression and Oncolysis in Combination with Low-Dose Cyclophosphamide Treatment on Antitumor Immune Responses
Cancer Res., January 15, 2006; 66(2): 960 - 969.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
C. Massa, C. Melani, and M. P. Colombo
Chaperon and Adjuvant Activity of hsp70: Different Natural Killer Requirement for Cross-Priming of Chaperoned and Bystander Antigens
Cancer Res., September 1, 2005; 65(17): 7942 - 7949.
[Abstract] [Full Text] [PDF]

Home page
 Neuro OncolHome page
M. W. Graner and D. D. Bigner
Chaperone proteins and brain tumors: Potential targets and possible therapeutics
Neuro-oncol, July 1, 2005; 7(3): 260 - 278.
[Abstract] [PDF]

Home page
 J. Virol.Home page
M. Kotsiopriftis, J. E. Tanner, and C. Alfieri
Heat Shock Protein 90 Expression in Epstein-Barr Virus-Infected B Cells Promotes {gamma}{delta} T-Cell Proliferation In Vitro
J. Virol., June 1, 2005; 79(11): 7255 - 7261.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. Demine and P. Walden
Testing the Role of gp96 as Peptide Chaperone in Antigen Processing
J. Biol. Chem., May 6, 2005; 280(18): 17573 - 17578.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
H. Zheng and Z. Li
Cutting Edge: Cross-Presentation of Cell-Associated Antigens to MHC Class I Molecule Is Regulated by a Major Transcription Factor for Heat Shock Proteins
J. Immunol., November 15, 2004; 173(10): 5929 - 5933.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
S.-J. Lee, L. Myers, G. Muralimohan, J. Dai, Y. Qiao, Z. Li, R. S. Mittler, and A. T. Vella
4-1BB and OX40 Dual Costimulation Synergistically Stimulate Primary Specific CD8 T Cells for Robust Effector Function
J. Immunol., September 1, 2004; 173(5): 3002 - 3012.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
D. SenGupta, P. J. Norris, T. J. Suscovich, M. Hassan-Zahraee, H. F. Moffett, A. Trocha, R. Draenert, P. J. R. Goulder, R. J. Binder, D. L. Levey, et al.
Heat Shock Protein-Mediated Cross-Presentation of Exogenous HIV Antigen on HLA Class I and Class II
J. Immunol., August 1, 2004; 173(3): 1987 - 1993.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
J. C. Baker-LePain, M. Sarzotti, and C. V. Nicchitta
Glucose-Regulated Protein 94/Glycoprotein 96 Elicits Bystander Activation of CD4+ T Cell Th1 Cytokine Production In Vivo
J. Immunol., April 1, 2004; 172(7): 4195 - 4203.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
C. Massa, C. Guiducci, I. Arioli, M. Parenza, M. P. Colombo, and C. Melani
Enhanced Efficacy of Tumor Cell Vaccines Transfected with Secretable hsp70
Cancer Res., February 15, 2004; 64(4): 1502 - 1508.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
M. P. Mycko, H. Cwiklinska, J. Szymanski, B. Szymanska, G. Kudla, L. Kilianek, A. Odyniec, C. F. Brosnan, and K. W. Selmaj
Inducible Heat Shock Protein 70 Promotes Myelin Autoantigen Presentation by the HLA Class II
J. Immunol., January 1, 2004; 172(1): 202 - 213.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
B. Liu, J. Dai, H. Zheng, D. Stoilova, S. Sun, and Z. Li
Cell surface expression of an endoplasmic reticulum resident heat shock protein gp96 triggers MyD88-dependent systemic autoimmune diseases
PNAS, December 23, 2003; 100(26): 15824 - 15829.
[Abstract] [Full Text] [PDF]

Home page
 J. Exp. Med.Home page
M. W. Hornef, B. H. Normark, A. Vandewalle, and S. Normark
Intracellular Recognition of Lipopolysaccharide by Toll-like Receptor 4 in Intestinal Epithelial Cells
J. Exp. Med., October 20, 2003; 198(8): 1225 - 1235.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
J. Stebbing, B. Gazzard, S. Portsmouth, F. Gotch, L. Kim, M. Bower, S. Mandalia, R. Binder, P. Srivastava, and S. Patterson
Disease-associated dendritic cells respond to disease-specific antigens through the common heat shock protein receptor
Blood, September 1, 2003; 102(5): 1806 - 1814.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
J. O. Jones and A. M. Arvin
Microarray Analysis of Host Cell Gene Transcription in Response to Varicella-Zoster Virus Infection of Human T Cells and Fibroblasts In Vitro and SCIDhu Skin Xenografts In Vivo
J. Virol., December 20, 2002; 77(2): 1268 - 1280.
[Abstract] [Full Text] [PDF]

Home page
 J. Exp. Med.Home page
J. C. Baker-LePain, M. Sarzotti, T. A. Fields, C.-Y. Li, and C. V. Nicchitta
GRP94 (gp96) and GRP94 N-Terminal Geldanamycin Binding Domain Elicit Tissue Nonrestricted Tumor Suppression
J. Exp. Med., December 2, 2002; 196(11): 1447 - 1459.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
K. Bethke, F. Staib, M. Distler, U. Schmitt, H. Jonuleit, A. H. Enk, P. R. Galle, and M. Heike
Different Efficiency of Heat Shock Proteins (HSP) to Activate Human Monocytes and Dendritic Cells: Superiority of HSP60
J. Immunol., December 1, 2002; 169(11): 6141 - 6148.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
E. Noessner, R. Gastpar, V. Milani, A. Brandl, P. J. S. Hutzler, M. C. Kuppner, M. Roos, E. Kremmer, A. Asea, S. K. Calderwood, et al.
Tumor-Derived Heat Shock Protein 70 Peptide Complexes Are Cross-Presented by Human Dendritic Cells
J. Immunol., November 15, 2002; 169(10): 5424 - 5432.
[Abstract] [Full Text] [PDF]

Home page
 Molecular Cancer TherapeuticsHome page
B. Liu, A. M. DeFilippo, and Z. Li
Overcoming Immune Tolerance to Cancer by Heat Shock Protein Vaccines
Mol. Cancer Ther., October 1, 2002; 1(12): 1147 - 1151.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. M. Vabulas, S. Braedel, N. Hilf, H. Singh-Jasuja, S. Herter, P. Ahmad-Nejad, C. J. Kirschning, C. da Costa, H.-G. Rammensee, H. Wagner, et al.
The Endoplasmic Reticulum-resident Heat Shock Protein Gp96 Activates Dendritic Cells via the Toll-like Receptor 2/4 Pathway
J. Biol. Chem., May 31, 2002; 277(23): 20847 - 20853.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
B. Berwin, J. P. Hart, S. V. Pizzo, and C. V. Nicchitta
CD91-Independent Cross-Presentation of GRP94(gp96)-Associated Peptides
J. Immunol., May 1, 2002; 168(9): 4282 - 4286.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zheng, H.
Right arrow Articles by Li, Z.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zheng, H.
Right arrow Articles by Li, Z.